Measurement of plasma renin activity by a simple solid phase radioimmunoassay.
A solid phase RIA in which an antibody adsorbed onto polystyrene balls was developed to determine PRA. Complete inhibition of converting enzyme and angiotensinase during enzymatic reaction was achieved by phenyl methyl sulfonyl fluoride and EDTA combination. Pepstatin A was found to be an effective agent to block angiotensin I generation during the RIA, and the sample can be directly incubated at room temperature for RIA without any special treatment to inhibit renin activity. The intra- and interassay coefficients of variation (CV) were 5.5-6.9% and 3.7-8.2%, respectively. The recovery was 91.9-117% of added angiotensin I. The assay is sufficiently sensitive and reliable for routine use and correlates acceptably (r = 0.996) with an established RIA. The antibody-adsorbed balls were compared to the soluble antibody with respect to thermodynamic parameters. It was found that the apparent equilibrium constant of the antibody-adsorbed ball was reduced to approximately 1/2 sec of soluble antibody, which was predominately due to the decrease in unitary entropy change by adsorption.
D000803:Angiotensin I / Q000276:immunology / Q000378:metabolism; D004492:Edetic Acid; D005665:Furosemide; D006801:Humans; D010436:Pepstatins; D010664:Phenylmethylsulfonyl Fluoride; D011137:Polystyrenes; D011863:Radioimmunoassay / Q000379:methods*; D012083:Renin / Q000097:blood*; D013816:Thermodynamics; D000092502:Renin Inhibitors
D000803:Angiotensin I; D004492:Edetic Acid; D005665:Furosemide; D010436:Pepstatins; D010664:Phenylmethylsulfonyl Fluoride; D011137:Polystyrenes; D012083:Renin; D000092502:Renin Inhibitors; C014195:Streptomyces pepsin inhibitor; C031375:pepstatin